The Human sTfR ELISA is a sandwich enzyme immunoassay for the quantitative measurement of human soluble transferrin receptor. In the Human sTfR ELISA, standards, quality controls and samples are incubated in microplate wells pre-coated with monoclonal anti-human sTfR antibody. After 60 minutesincubation and washing, monoclonal anti-human sTfR antibody, conjugated with horseradish peroxidase (HRP) is added to the wells and incubated for 60 minutes with captured sTfR. Following another washing step, the remaining HRP conjugate is allowed to react with the substrate solution (TMB). The reaction is stopped by addition of acidic solution and absorbance of the resulting yellow product is measured. The absorbance is proportional to the concentration of sTfR. A standard curve is constructed by plotting absorbance values against concentrations of standards, and concentrations of unknown samples are determined using this standard curve.